Ion [35]. The MDA content material at 532 nm was calculated by subtracting the absorbance at 600 nm. 2.5. Leaf Photosynthesis, Chlorophyll Fluorescence Parameters, and Chlorophyll Content material The net photosynthetic rate (Pn), stomatal conductance (Gs), transpiration rate (Tr), and intercellular CO2 concentration (Ci) on the leaves had been measured by the portable photosynthetic system (li-6400, Li-COR, Lincoln, NE, USA). Leaf photosynthetic parameters were determined at 10 a.m. following the plants have been treated with distinctive concentrations of NaCl and treated with distinctive concentrations of calcium chloride for a single week. The mature leaves have been dark-adapted for 20 min devoid of isolation, and the fluorescence kinetic parameters at room temperature were measured using a transportable modulation chlorophyll fluorescence instrument (PAM-2500 Walz, Effeltrich, Germany). For the chlorophyll content material, 0.03 g of fresh leaves have been extracted inside a 10 mL pigment extraction solution containing absolute ethanol and acetone (1:2, v/v) at 25 C for 12 h in the dark. The absorbance of the supernatant at 470, 645, and 663 nm was then measured making use of an ultraviolet spectrophotometer. Chlorophyll a, chlorophyll b, carotenoids, and total chlorophyll content were calculated based on [36]. two.six. Determination of K+ , Na+ , and Ca2+ To figure out the K+ , Na+ , and Ca2+ ion concentrations, we cautiously washed fresh root, stem, and leaf samples with deionized water, placed them in an oven at 105 C for 20 min, after which kept the temperature constant at 80 C till the samples were fully dried. The dried plant samples were then grounded inside a 5 mL centrifuge tubes utilizing a high-throughput plant Semicarbazide (hydrochloride) supplier tissue ball milling instrument (Scientz-192, Xinzhi Biotechnology Co., Ltd., Ningbo, China). A total of 0.3 g of every sample powder was weighed, and 5 mL of nitric acid and 1 mL of perchloric acid had been added for wet digestion. The K+ , Na+ , and Ca2+ contents of plant tissue extracts and regular samples (National Institute of Metrology, Beijing, China) had been determined by inductively coupled plasma optical emission spectrometer (ICP-OES; PerkinElmer, Optima 8300, Waltham, MA, USA). The concentration of K+ , Na+ , and Ca2+ is defined as K+ , Na+ , and Ca2+ content material (mg) per unit tissue (g) [37]. two.7. Extraction and LC S Evaluation of Phenolic Compounds 2.7.1. Chemical substances and Reagents UPLC-grade acetonitrile and methanol had been bought from Fisher Scientific (Pittsburgh, PA, USA). All other reagents have been of analytical purity. Ultrapure water was prepared by a Milli-Q system (Millipore, Bedford, MA, USA) water purification program. The reference compounds essential for the experiment were all purchased from ChromaDex Inc. (Santa Ana, CA, USA), like p-hydroxycinnamic acid, p-hydroxybenzoic acid, two,5-dihydroxybenzoic acid, genistein, abscisic acid, petunidin, naringenin, hesperidin, quercetin-3-O-rhamnoside, chlorogenic acid, ferulic acid, myricetin, Trilinolein Cancer luteolin, catechin, cinnamic acid, p-coumaric acid, hesperetin, quercetin, caffeic acid, L-phenylalanine, naringin, kaempferol, liquiritigenin, isoliquiritigenin, and vanillic acid. The purities of those standards were higher than 98 .Agriculture 2021, 11,5 of2.7.2. Preparation of Test Sample Solution Gleditsia sinensis plant tissues (root, stem, and leaf) treated with various remedies (CK, S1, S2, S1 + C1, S1 + C2, S1 + C3) had been grounded and then ultrasonically extracted (100 kHz, 40) for 45 min by adding 10 mL of 70 methanol. Following filtration, the.
